作者没有致谢。

所报告的方法被用于一项研究, 它得到了伯恩茅斯大学研究伦理学委员会2016年2月9日的伦理批准 (参考文献 10571). 
 1. LSCI 设置 <ol> <li> 将 moorFLPI LSCI 后面板连接到其三插槽 (电源、通用串行总线 (USB) 和 IEEE1394), 以便系统正常工作. </li>
	<li> 使用4螺钉将桌面支持臂装配到 moorFLPI LSCI 翻转, 并固定在支撑臂上. </li>
	<li> 旋转安装托架以便在连接时向下成像. 
	注: LSCI 有三控制: (1) 变焦调节器和 #8211; 随着成像器设置在一个位置, 较小的放大倍数可以分别调整为较小和更大的视野的最大和最小设置。为确保重复性, 提供了一个索引环标签;(2) 焦点调节器和 #8211; 这取决于测量距离, 必须在设置图像位置后进行调整。为确保重复性, 提供了一个索引环标签;和 (3) 偏光片和 #8211; 线性偏振滤波器可使曝光的机构和 #8211 的镜面反射最小化; 旋转安装可通过360和 #176 进行翻转;. </li>
	<li> 安装软件来控制摄像头。该软件分为两个模块, 提供了一个测量和审查功能. </li>
</ol> 2。参与者准备 <ol> <li> 确保在温度控制室 (22 和 #177; 1 和 #176; C) 中执行评估, 并且参与者在测试前10分钟就位, 以适应室温. </li>
	<li> 避免强烈的人工光源和阳光照射在参与者或 LSCI 上, 因为环境光线会影响 moorFLPI 近红外激光源的运行在 785 nm. 
	注: 一个简单的测试, 以确定是否可以接受的环境光线水平是通过打开成像设置窗口和阻碍激光。如果图像是几乎完全黑色的, 那么不需要进一步的步骤;如果仍然存在太多的环境光, 则需要进一步的操作. </li>
	<li> 确保参与者在整个评估过程中都能轻松自如, 如果坐在地上, 并避免对话. </li>
	<li> 放置 8 cm 2 粘附不透明贴片 (AOP) (如 、Leukotape) 在皮肤区域上以遮盖血流。这是为了解释 LSCI 的缺点, 在高灵敏度的工件运动, 和信号后向散射将用于测量微循环血流. </li>
</ol> 3。微循环图像测量 <ol> <li> 选择和 #39; 空间处理和 #39; 每秒捕获25帧, 以 152 x 113 像素为单位. </li>
	<li> 选择和 #39; 实时图像测量和 #39; 调整成像器20厘米以外的位置, 然后调整缩放、焦点和偏光镜以最小的镜面反射。图像应该显得相当和 #39; 平面和 #39; 和无特征. </li>
	<li> 设置20毫秒的曝光时间, 以对小的变化和低通量进行高灵敏度. </li>
	<li> 使用25赫兹的显示速率和0.3 秒的时间常数来解释快速的血流变化, 并通过降低图像噪声来达到最佳对比度. </li>
	<li> 创建两个相等的大小 (2 cm 2 ) 感兴趣的区域 (roi), 称为 roi 1 和 roi 2。使 ROI 2 对齐, 因此它位于 8 cm 2 AOP 中。小心, 使 roi 不交换, 但保持接近2-4 厘米, 以减少 re-centering 的需要, 如果任何机械运动导致 ROI 2 不再在 AO 地区. 
	注意: 在低强度和高强度的区域, 血流测量将不太准确, 所以有一个最佳增益设置是很重要的。增益值范围介于 0-200 之间。在 70-80 范围内实现了最佳增益设定值. </li>
	<li> 相对于参照基线执行流量测量。在这种方法的情况下, 引入和 #39; 休息和 #39; 阶段作为参考基线。因此, 表达 #39; 快速和 #39; 和 #39; 慢速和 #39; 从基线、#39、休息和 #39, 阶段的血流百分比变化; 阶段. </li>
	<li> 以视频格式记录血流测量, 并使用图像复查模块保存离线分析. </li>
</ol> 4。脱机分析 注意: moorFLPI 图像审阅软件允许打开视频进行分析. 
<ol> <li> 在一系列的平均血流记录后, 计算 roi 内的平均通量。roi 1 是对皮肤血流的真实测量, roi 2 是来自 AOP 的散射激光散斑皮肤信号. </li>
	<li> 通过从 roi 1 (皮肤血流) 中减去 roi 2 来计算平均血流. 
	roi 1-roi 2 = 平均血流 </li> </ol>

<ol>
	<li>Mahe, G., Rousseau, P., Durand, S., Leftheriotis, G., Abraham, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+speckle+contract+imaging+accurately+measures+blood+flow+over+moving+skin+surfaces.">Laser speckle contract imaging accurately measures blood flow over moving skin surfaces.</a> Microvas Res. 81 (2), 183-188 (2010).</li><li>Rousseau, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increasing+the+&amp;#34;region+of+interest&amp;#34;+and+&amp;#34;time+of+interest&amp;#34;,+both+reduce+the+variability+of+blood+flow+measurements+using+laser+speckle+contrast+imaging.">Increasing the &amp;#34;region of interest&amp;#34; and &amp;#34;time of interest&amp;#34;, both reduce the variability of blood flow measurements using laser speckle contrast imaging.</a> Microvas Res. 82, 88-91 (2011).</li><li>Hecht, N., Woitzik, J., Dreier, J., Vajkoczy, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+monitoring+of+cerebral+blood+flow+by+laser+speckle+contrast+analysis.">Intraoperative monitoring of cerebral blood flow by laser speckle contrast analysis.</a> Neurosurg Focus. 27 (4), 1-6 (2009).</li><li>Mahe, G., Durand, S., Humeau-Heurtier, A., Leftheriotis, G., Abrham, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+experimental+conditions+on+noncontact+laser+recordings+in+microvascular+studies.">Impact of experimental conditions on noncontact laser recordings in microvascular studies.</a> Microcirculation. 19 (8), 669-675 (2012).</li><li>Cheng, H., Duong, T. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+laser-speckle-imaging+analysis+method+and+its+application+to+retinal+blood+flow+imaging.">Simplified laser-speckle-imaging analysis method and its application to retinal blood flow imaging.</a> Opt Lett. 32 (15), 2188-2190 (2007).</li><li>Doherty, J., McNamara, P., Clancy, N. T., Enfield, J. G., Leahy, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+instruments+for+investigation+of+microcirculatory+blood+flow+and+red+blood+cell+concentration.">Comparison of instruments for investigation of microcirculatory blood flow and red blood cell concentration.</a> J Biomed Opt. 14 (3), 034025 (2009).</li><li>Briers, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+speckle+contrast+imaging+for+measuring+blood+flow.">Laser speckle contrast imaging for measuring blood flow.</a> Opt Appl. 37 (1), 139-152 (2007).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> moorFLPI User Manual. (8), (2012).</li><li>Mahe, G., Durand, S., Humeau-Heurtier, A., Abraham, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+Experimental+Conditions+on+Noncontact+Laser+Recordings+in+Microvascular+Studies.">Impact of Experimental Conditions on Noncontact Laser Recordings in Microvascular Studies.</a> Microcirculation. 19, 669-675 (2012).</li><li>Bahadori, S., Immins, T., Wainwright, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+calf+neuromuscular+electrical+stimulation+and+intermittent+pneumatic+compression+on+thigh+microcirculation.">The effect of calf neuromuscular electrical stimulation and intermittent pneumatic compression on thigh microcirculation.</a> Micro Res. 111, 37-41 (2017).</li><li>Omarjee, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimisation+of+movement+detection+and+artifact+removal+during+laser+speckle+contrast+imaging.">Optimisation of movement detection and artifact removal during laser speckle contrast imaging.</a> Miscrovas Res. 97 (1), 75-80 (2015).</li></ol>

作者没有什么可透露的。

本研究的目的是介绍一种新的方法来测量和分析的交替速度的血液微循环在一个单一的实验使用 LSCI。测量可以受到环境光、振动和参与运动的影响, 包括呼吸和抽搐。该协议中概述的步骤都是为了尽量减少这些影响, 并获得可靠和可重复的血液微循环测量。
人们强调, 该协议中的每一个步骤对于准确测量血液微循环都是至关重要的, 在对所有可能的设置选项和组合进行顺序测试后发现了该技术, 包括:时间常数, 变焦设置, 曝光时间, 显示率, 增益和通量图像调色板。结果进行了分析和重复使用现场视频显示和离线分析找到最佳的成像设置。这是必不可少的, 因为图像处理软件使用的事实, 高灌注产生的激光散斑模式的快速变化, 并因此在视频图像中产生了一个低对比度清晰的散斑区。灌注图像, 然后创建在一个 color-coded 的微循环灌注图。
实验区和参与准备是必不可少的, 这可以通过避免工作附近的日光源 (窗口) 或强光源的人工光, 因为这些可能会干扰 moorFLPI 附近的红外激光源。该协议还介绍了一个 AOP, 因为它被承认, 环境振动和参与者的运动都产生的信号是无法区分的血流。AOP 证明是一个简单而有效的选择, 提供了一个薄但不透明, 光和容易接近的选项, 有一个微观粗糙的表面积, 以避免显着镜面反射。Omarjee et al.的初步研究11突出显示了一个潜在的限制, 即 Leukotape 创建的反射信号振幅与皮肤不同, 并且各主体之间的差异很大;然而马希et al.1发现参与者之间没有显著差异。虽然 Leukotape 比其他定制的双层粘接片更易访问, 但通过使用另一种 AOP, 可以提高方程 (1) 测量的精确度。
离线分析部分强调了 roi 大小的重要性, 以及它们在感兴趣区域的位置。最初, 一个更大的 roi 1 (大约 8 cm2) 被尝试, 其中覆盖了 roi 2。这种方法很快成为 un-reliable 由于工件运动导致的 roi 2 移动和现场实验必须停止, 以重新的 roi 2。另外一个短期的原因是, 由于 ROI 1 覆盖了 aop, 平均通量不再将 aop 下的区域作为一个帐户, 因为不再有散射信号。这意味着大量的血液微循环被忽视, 因此产生的通量数据是不正确的。因此, 一种方法, 其中两个 roi 2 cm2, 具有 8 cm2的 AOP, 在 roi 1 和 roi 2 之间没有交互 (但在彼此之间保持2-4 厘米), 为血液测量提供了可靠且可重复的分析技术。微循环.

激光散斑造影成像 (LSCI) 是一种行之有效的非接触、实时监测方法, 用于分析血流微循环1,2,3,45,6,7. 本文所用的 LSCI moorFLPI 全域。利用一种称为 "激光散斑" 的现象测量大面积高时空分辨率的血流灌注是该装置6的主要优点之一。real-time 评估的微循环是通过拍摄的模式, 通过相机使用散斑模式。鉴于 moorFLPI LSCI 用于临床和生理学研究, 图像处理软件的工作原理是, 高灌注产生的激光散斑模式的快速变化, 然后由带电耦合器件 (CCD) 集成生成低对比度的区域8。对比度是量化的, 由此产生的流量是彩色编码产生一个灌注图像8。
不幸的是, LSCI 是非常敏感的环境振动, 工件和运动的主题领域9。到目前为止, 这为交流流状态的研究提供了挑战。本文详细介绍了显式技术概述了在最近的研究10 , 其中一个神经肌肉电刺激装置被用来测量血液微循环时, 有肢体的运动正在检查。

<table border="0" cellpadding="0" cellspacing="0" style="width:1117px;" width="1117">
	<tbody>
		<tr height="64">
			<td height="64" style="height:64px;width:260px;">moorFLPI LSCI</td>
			<td style="width:188px;">Moor Instruments</td>
			<td style="width:152px;">Not Available - Online Link Provided in descreption</td>
			<td style="width:517px;">moorFLPI is an instrument designed for the measurment of blood flow within microvasculature by using infra red laser speckle contrast analysis.&#160; https://gb.moor.co.uk/</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:260px;">moorFLPI Image Review Module</td>
			<td style="width:188px;">Moor Instruments</td>
			<td style="width:152px;">No Available - Online Link Provided</td>
			<td style="width:517px;">Used with moorFLPI, user can record and measure changes in blood flow by changerating a colour coded map of tissue perfusion.&#160; https://gb.moor.co.uk/</td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:260px;">Leukotape</td>
			<td style="width:188px;">BSN Medical</td>
			<td style="width:152px;">72978-10</td>
			<td style="width:517px;">Medical tape with microporous surface. http://www.bsnmedical.co.uk/fileadmin/z-countries/United_Kingdom/PDF/L/Leukotape_K_A46PP_low_res_11112013.pdf</td>
		</tr>
	</tbody>
</table>

a novel approach to overcome movement artifact when using a laser speckle contrast imaging system for alternating speeds of blood microcirculation

激光散斑造影成像仪 (LSCI) 提供了一种强大而简单的测量微循环血流的技术。理想的血液动力反应, LSCI 的使用方式与传统的激光多普勒成像仪 (LDI) 相同。然而, 最大的皮肤深度约1毫米, LSCI 设计的重点主要是表面血流。它用于测量 15 cm x 20 cm 的皮肤表面积。本文介绍的新技术是微循环的交替速度;即使用 LSCI 的慢速和快速流量测量。该技术还克服了 LSCI 最大的缺点, 即对工件运动的灵敏度高。通过从激光散斑皮肤信号中减去来自 aop 的 LSCI 信号, 引入了一种粘着不透明贴片 (aop) 来满足微循环血流的满意记录。最佳的设置也被定义了, 因为 LSCI 是最强有力的, 当通量变动测量相对于参考基线, 以血液微循环通量表示作为一个百分比变动从基线。这些变化可用于分析血流系统的状态。

在图 1中概述了 LSCI 实验设置, 并确定了功能工具。一个典型的参与者准备测量的血流量在前大腿区域的说明。可调式安装支架允许旋转 LSCI 的血液流量测量在任何特定的皮肤区域微血管。图 2概述了典型的原始散斑图像和转换的散斑图像的示例, 并在《血液微循环测量协议》中列出了定制的设置。
图 3显示了 AOP 的皮肤区域和位置的示例 (步骤 3.1), LSCI 原始成像设置 (步骤 3.2), 一个慢血流的实时图像 (步骤 3.3) 和一个快速血流的实时图像 (步骤 3.4) 实现了在连续的数据记录的交替通量美国moorFLPI LSCI调色板 color-coding 设置允许差异之间的流量水平。与标准调色板的16种颜色, 低通量被视为蓝色, 中通量值被视为绿色和高通量值被视为橙色和红色。
皮肤血流是用激光散斑灌注单元 (LSPU) 来表达的。图 4显示了一个皮肤区域的 roi 1、roi 2 和 AOP 的图形表示形式。在离线分析中, 使用 roi 1 和 roi 2 的数据和公式 (1) 计算平均血流。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56415/56415fig1.jpg"> 
图 1: moorFLPI LSCI 实验设置.台式机支持臂、电缆输出、位置控制 (变焦调节器、聚焦调节器和偏振器)、AOP 和膝上型计算机用于配置图像设置。<a href="//ecsource.jove.com/files/ftp_upload/56415/56415fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56415/56415fig2.jpg"> 
图 2: 数据记录前的原始散斑图像2.1-2.2) 流量和原始散斑图像的配置糟糕, 导致高增益, 能见度差, 这将导致一个不准确的血液流量测量。2.3-2.4) 系统配置为按协议, 导致正确的增益和最大的能见度可靠的结果。<a href="//ecsource.jove.com/files/ftp_upload/56415/56415fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56415/56415fig3.jpg"> 
图 3: 使用 moorFLPI LSCI 设置和记录测量的概述.3.1. 具有 2 cm2 AOP 的皮肤 (大腿) 区域, 用于计算工件移动。3.2) 一个原始散斑 ' 平面和无特征 ' 图像表明良好的背向散射光强的最佳设置。3.3) 血流缓慢的实时图像记录。3.4) 一个快速血流的实时图像记录。<a href="//ecsource.jove.com/files/ftp_upload/56415/56415fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56415/56415fig4.jpg"> 
图 4: roi 1 的图形表示、roi 2 和在皮肤区域上的 AOP 布局.<a href="//ecsource.jove.com/files/ftp_upload/56415/56415fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td colspan="3">休息 (基线参考) (LSPU)</td>
			<td colspan="4">慢血流量 (LSPU)</td>
			<td colspan="4">适度血流 (LSPU)</td>
			<td colspan="4">快速血流 (LSPU)</td>
		</tr>
		<tr>
			<td>平均通量-ROI 1</td>
			<td>平均通量-ROI 2</td>
			<td>平均血流</td>
			<td>平均通量-ROI 1</td>
			<td>平均通量-ROI 2</td>
			<td>平均血流</td>
			<td>血流量从基线增加%</td>
			<td>平均通量-ROI 1</td>
			<td>平均通量-ROI 2</td>
			<td>平均血流</td>
			<td>血流量从基线增加%</td>
			<td>平均通量-ROI 1</td>
			<td>平均通量-ROI 2</td>
			<td>平均血流</td>
			<td>血流量从基线增加%</td>
		</tr>
		<tr>
			<td>157。9</td>
			<td>35。1</td>
			<td>122。8</td>
			<td>178。5</td>
			<td>41。6</td>
			<td>136。9</td>
			<td>10。9</td>
			<td>216。9</td>
			<td>44。6</td>
			<td>172。3</td>
			<td>33。5</td>
			<td>418。9</td>
			<td>77。5</td>
			<td>341。4</td>
			<td>94。2</td>
		</tr>
	</tbody>
</table>
表 1: LSPU 中的平均通量, 用于 roi 1 和 roi 2 的基线、慢速、中度和快速血流.血流的增加是表示从基线阶段的百分比变化。
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56415/56415fig5.jpg"> 
图 5: roi 1 和 roi 2 在皮肤表面积 (大腿) 上定位的示例.16色调色板概述了显影的克里斯·赖维斯。数据记录的 roi 1 和 roi 2 在灌注单元和减去如解释的公式1的血液微循环的测量。<a href="//ecsource.jove.com/files/ftp_upload/56415/56415fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about A Novel Approach to Overcome Movement Artifact When Using a Laser Speckle Contrast Imaging System for Alternating Speeds of Blood Microcirculation at JoVE.com

A Novel Approach to Overcome Movement Artifact When Using a Laser Speckle Contrast Imaging System for Alternating Speeds of Blood Microcirculation

用激光散斑对比成像系统克服运动伪影的新方法血液微循环的交替速度

本研究介绍了一种利用激光散斑造影仪测量和分析血液微循环交替速度的新方法。

The laser speckle contrast imager (LSCI) provides a powerful yet simple technique for measuring microcirculatory blood flow. Ideal for blood dynamic ...

a-novel-approach-to-overcome-movement-artifact-when-using-laser

Research

JoVE Journal

Medicine

8.3K 次观看.  Bournemouth University. 本研究介绍了一种利用激光散斑造影仪测量和分析血液微循环交替速度的新方法。

视频: A Novel Approach to Overcome Movement Artifact When Using a Laser Speckle Contrast Imaging System for Alternating Speeds of Blood Microcirculation

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading to an accumulation of lipid-laden intra-luminal plaque, smooth muscle cell proliferation and progressive narrowing of the vessel lumen. The formation of such vulnerable plaques prone to rupture underlies the majority of cases of acute myocardial infarction. The complex molecular and cellular inflammatory cascade is orchestrated by the recruitment of T lymphocytes and macrophages and their paracrine effects on endothelial and smooth muscle cells.1
Molecular imaging in atherosclerosis has evolved into an important clinical and research tool that allows in vivo visualization of inflammation and other biological processes. Several recent examples demonstrate the ability to detect high-risk plaques in patients, and assess the effects of pharmacotherapeutics in atherosclerosis.4 While a number of molecular imaging approaches (in particular MRI and PET) can image biological aspects of large vessels such as the carotid arteries, scant options exist for imaging of coronary arteries.2 The advent of high-resolution optical imaging strategies, in particular near-infrared fluorescence (NIRF), coupled with activatable fluorescent probes, have enhanced sensitivity and led to the development of new intravascular strategies to improve biological imaging of human coronary atherosclerosis.
Near infrared fluorescence (NIRF) molecular imaging utilizes excitation light with a defined band width (650-900 nm) as a source of photons that, when delivered to an optical contrast agent or fluorescent probe, emits fluorescence in the NIR window that can be detected using an appropriate emission filter and a high sensitivity charge-coupled camera. As opposed to visible light, NIR light penetrates deeply into tissue, is markedly less attenuated by endogenous photon absorbers such as hemoglobin, lipid and water, and enables high target-to-background ratios due to reduced autofluorescence in the NIR window. Imaging within the NIR 'window' can substantially improve the potential for in vivo imaging.2,5
Inflammatory cysteine proteases have been well studied using activatable NIRF probes10, and play important roles in atherogenesis. Via degradation of the extracellular matrix, cysteine proteases contribute importantly to the progression and complications of atherosclerosis8. In particular, the cysteine protease, cathepsin B, is highly expressed and colocalizes with macrophages in experimental murine, rabbit, and human atheromata.3,6,7 In addition, cathepsin B activity in plaques can be sensed in vivo utilizing a previously described 1-D intravascular near-infrared fluorescence technology6, in conjunction with an injectable nanosensor agent that consists of a poly-lysine polymer backbone derivatized with multiple NIR fluorochromes (VM110/Prosense750, ex/em 750/780nm, VisEn Medical, Woburn, MA) that results in strong intramolecular quenching at baseline.10 Following targeted enzymatic cleavage by cysteine proteases such as cathepsin B (known to colocalize with plaque macrophages), the fluorochromes separate, resulting in substantial amplification of the NIRF signal. Intravascular detection of NIR fluorescence signal by the utilized novel 2D intravascular NIRF catheter now enables high-resolution, geometrically accurate in vivo detection of cathepsin B activity in inflamed plaque.
In vivo molecular imaging of atherosclerosis using catheter-based 2D NIRF imaging, as opposed to a prior 1-D spectroscopic approach,6 is a novel and promising tool that utilizes augmented protease activity in macrophage-rich plaque to detect vascular inflammation.11,12 The following research protocol describes the use of an intravascular 2-dimensional NIRF catheter to image and characterize plaque structure utilizing key aspects of plaque biology. It is a translatable platform that when integrated with existing clinical imaging technologies including angiography and intravascular ultrasound (IVUS), offers a unique and novel integrated multimodal molecular imaging technique that distinguishes inflammatory atheromata, and allows detection of intravascular NIRF signals in human-sized coronary arteries.

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.

体内近红外荧光（NIRF）血管内分子成像炎性斑块，多式联运方针成像动脉粥样硬化

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

科研

医学

Contrast Enhanced Vessel Imaging using MicroCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between different soft tissues and body fluids is inherently poor in micro-CT images 1. Under these circumstances, visualization of blood vessels becomes a nearly impossible task. To overcome this and to improve the visualization of blood vessels exogenous contrast agents can be used. Herein, we present a methodology for visualizing the vascular network in a rodent model. By using a long-acting aqueous colloidal polydisperse iodinated blood-pool contrast agent, eXIA 160XL, we optimized image acquisition parameters and volume-rendering techniques for finding blood vessels in live animals. Our findings suggest that, to achieve a superior contrast between bone and soft tissue from vessel, multiple-frames (at least 5-8/ frames per view), and 360-720 views (for a full 360&deg; rotation) acquisitions were mandatory. We have also demonstrated the use of a two-dimensional transfer function (where voxel color and opacity was assigned in proportion to CT value and gradient magnitude), in visualizing the anatomy and highlighting the structure of interest, the blood vessel network. This promising work lays a foundation for the qualitative and quantitative assessment of anti-angiogenesis preclinical studies using transgenic or xenograft tumor-bearing mice.

Contrast enhanced small animal vessel imaging by microCT is a rapid, cost-effective and high-throughput technique for serial in situ examination for tumor development, for analyzing the network of blood vessels that nourish them, and for following the response of tumors to preclinical therapeutic intervention(s).

增强容器成像MicroCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between ...

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks. Bronchial thermoplasty is delivered by the Alair System and is performed in three outpatient procedure visits, each scheduled approximately three weeks apart. The first procedure treats the airways of the right lower lobe, the second treats the airways of the left lower lobe and the third and final procedure treats the airways in both upper lobes. After all three procedures are performed the bronchial thermoplasty treatment is complete.
Bronchial thermoplasty is performed during bronchoscopy with the patient under moderate sedation. All accessible airways distal to the mainstem bronchi between 3 and 10 mm in diameter, with the exception of the right middle lobe, are treated under bronchoscopic visualization. Contiguous and non-overlapping activations of the device are used, moving from distal to proximal along the length of the airway, and systematically from airway to airway as described previously. Although conceptually straightforward, the actual execution of bronchial thermoplasty is quite intricate and procedural duration for the treatment of a single lobe is often substantially longer than encountered during routine bronchoscopy. As such, bronchial thermoplasty should be considered a complex interventional bronchoscopy and is intended for the experienced bronchoscopist. Optimal patient management is critical in any such complex and longer duration bronchoscopic procedure. This article discusses the importance of careful patient selection, patient preparation, patient management, procedure duration, postoperative care and follow-up to ensure that bronchial thermoplasty is performed safely.
Bronchial thermoplasty is expected to complement asthma maintenance medications by providing long-lasting asthma control and improving asthma-related quality of life of patients with severe asthma. In addition, bronchial thermoplasty has been demonstrated to reduce severe exacerbations (asthma attacks) emergency rooms visits for respiratory symptoms, and time lost from work, school and other daily activities due to asthma.

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks.

支气管Thermoplasty:严重哮喘的一种新的治疗方法

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

脓毒症动物模型，用于评估新型草药疗法

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

Intraoperative Detection of Subtle Endometriosis: A Novel Paradigm for Detection and Treatment of Pelvic Pain Associated with the Loss of Peritoneal Integrity

Endometriosis is a common disease affecting 40 to 70% of reproductive-aged women with chronic pelvic pain (CPP) and/or infertility. The purpose of this study was to demonstrate the use of a blue dye (methylene blue) to stain peritoneal surfaces during laparoscopy (L/S) to detect the loss of peritoneal integrity in patients with pelvic pain and suspected endometriosis. Forty women with CPP and 5 women without pain were evaluated in this pilot study. During L/S, concentrated dye was sprayed onto peritoneal surfaces, then aspirated and rinsed with Lactated Ringers solution. Areas of localized dye uptake were evaluated for the presence of visible endometriotic lesions. Areas of intense peritoneal staining were resected and some fixed in 2.5% buffered gluteraldehyde and examined by scanning (SEM) electron microscopy. Blue dye uptake was more common in women with endometriosis and chronic pelvic pain than controls (85% vs. 40%). Resection of the blue stained areas revealed endometriosis by SEM and loss of peritoneal cell-cell contact compared to normal, non-staining peritoneum. Affected peritoneum was associated with visible endometriotic implants in most but not all patients. Subjective pain relief was reported in 80% of subjects. Based on scanning electron microscopy, we conclude that endometrial cells extend well beyond visible implants of endometriosis and appear to disrupt the underlying mesothelium. Subtle lesions of endometriosis could therefore cause pelvic pain by disruption of peritoneal integrity, allowing menstrual or ovulatory blood and associated pain factors access to underlying sensory nerves. Complete resection of affected peritoneum may provide a better long-term treatment for endometriosis and CPP. This simple technique appears to improve detection of subtle or near invisible endometriosis in women with CPP and minimal visual findings at L/S and may serve to elevate diagnostic accuracy for endometriosis at laparoscopy.

Loss of peritoneal integrity provides a new paradigm to understand and treat chronic pelvic pain in women with mild forms of endometriosis and can be easily detected using intraoperative instillation of dye at the time of laparoscopy.

术中的微妙的子宫内膜异位症的检测:A新型范式为相关联的与的的腹膜的完整性之亏损盆腔疼痛的的的的检测和治疗的

Endometriosis is a  common disease affecting 40 to 70% of reproductive-aged women with chronic  pelvic pain (CPP) and/or infertility. The purpose of this ...

Measurement of Dynamic Scapular Kinematics Using an Acromion Marker Cluster to Minimize Skin Movement Artifact

The measurement of dynamic scapular kinematics is complex due to the sliding nature of the scapula beneath the skin surface. The aim of the study was to clearly describe the acromion marker cluster (AMC) method of determining scapular kinematics when using a passive marker motion capture system, with consideration for the sources of error which could affect the validity and reliability of measurements. The AMC method involves placing a cluster of markers over the posterior acromion, and through calibration of anatomical landmarks with respect to the marker cluster it is possible to obtain valid measurements of scapular kinematics. The reliability of the method was examined between two days in a group of 15 healthy individuals (aged 19-38 years, eight males) as they performed arm elevation, to 120&#176;, and lowering in the frontal, scapular and sagittal planes. Results showed that between-day reliability was good for upward scapular rotation (Coefficient of Multiple Correlation; CMC = 0.92) and posterior tilt (CMC = 0.70) but fair for internal rotation (CMC = 0.53) during the arm elevation phase. The waveform error was lower for upward rotation (2.7&#176; to 4.4&#176;) and posterior tilt (1.3&#176; to 2.8&#176;), compared to internal rotation (5.4&#176; to 7.3&#176;). The reliability during the lowering phase was comparable to results observed during the elevation phase. If the protocol outlined in this study is adhered to, the AMC provides a reliable measurement of upward rotation and posterior tilt during the elevation and lowering phases of arm movement.

This report presents details of how to adopt the acromion marker cluster method of obtaining scapular kinematics when using a passive marker motion capture device. As has been described in the literature, this method provides a robust, non-invasive, three-dimensional, dynamic and valid measurement of scapular kinematics, minimizing skin movement artifact.

动态肩胛运动学使用的肩峰标记集群尽量减少与皮肤运动伪影测量

The measurement of dynamic scapular kinematics is complex due to the sliding nature of the scapula beneath the skin surface. The aim of the study was to ...

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 &#181;l) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described &#8220;ischemic penumbra zone&#8221;. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.

Contrast Enhanced Ultrasound imaging is a reliable in-vivo tool for quantifying spinal cord blood flow in an experimental rat spinal cord injury model. This paper contains a comprehensive protocol for application of this technique in association with a contusion model of thoracic spinal cord injury.

超声造影成像脊髓血流量的评估实验脊髓损伤

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an ...

3D Ultrasound Imaging: Fast and Cost-effective Morphometry of Musculoskeletal Tissue

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS images are constructed from calibrated freehand 2D B-mode ultrasound images, which are positioned into a voxel array. Ultrasound (US) imaging allows quantification of muscle size, fascicle length, and angle of pennation. These morphological variables are important determinants of muscle force and length range of force exertion. The presented protocol describes an approach to determine volume and fascicle length of m.&#160;vastus lateralis and m.&#160;gastrocnemius medialis. 3DUS facilitates standardization using 3D anatomical references. This approach provides a fast and cost-effective approach for quantifying 3D morphology in skeletal muscles. In healthcare and sports, information on the morphometry of muscles is very valuable in diagnostics and/or follow-up evaluations after treatment or training.

3D ultrasound imaging (3DUS) allows fast and cost-effective morphometry of musculoskeletal tissues. We present a protocol to measure muscle volume and fascicle length using 3DUS.

3D 超声成像: 快速、经济高效的肌骨骼组织形态分析

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

小鼠体外循环模型: 一种新方法

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

As a functional status of microcirculation, microvascular vasomotion is important for the delivery of oxygen and nutrients and the removal of carbon dioxide and waste products. The impairment of microvascular vasomotion might be a crucial step in the development of microcirculation-related diseases. In addition, the highly vascularized pancreatic islet is adapted to support endocrine function. In this respect, it seems possible to infer that the functional status of pancreatic islet microvascular vasomotion might affect pancreatic islet function. Analyzing the pathological changes of the functional status of pancreatic islet microvascular vasomotion may be a feasible strategy to determine contributions that pancreatic islet microcirculation makes to related diseases, such as diabetes mellitus, pancreatitis, etc. Therefore, this protocol describes using a laser Doppler blood flow monitor to determine the functional status of pancreatic islet microvascular vasomotion, and to establish parameters (including average blood perfusion, amplitude, frequency, and relative velocity of pancreatic islet microvascular vasomotion) for evaluation of the microcirculatory functional status. In a streptozotocin-induced diabetic mouse model, we observed an impaired functional status of pancreatic islet microvascular vasomotion. In conclusion, this approach for assessing pancreatic islet microvascular vasomotion in vivo may reveal mechanisms relating to pancreatic islet diseases.